Fig. 6.
Overexpression of SQSTM1 increases misfolded SOD1 in the spinal cord of SOD1H46R-expressing mice. a Western blot analysis of SOD1. b Western blot analysis of misfolded SOD1. a, b The spinal cord samples were obtained from wild-type (WT), SQSTM1 (SQSTM1), SOD1H46R (H46R), and SQSTM1;SOD1H46R (SQSTM1;H46R) mice at 16 weeks of age (wk), 22 wk., and end-stage (H46R and SQSTM1;H46R) or 28 wk. (WT and SQSTM1). Two fractions; 1% Triton X-100 soluble and 1% Triton X-100 insoluble/5%SDS soluble fractions were analyzed. SOD1_mono and SOD1_HMW represent monomeric and high molecular-weight forms of SOD1, respectively. SOD1(C4F6) represents misfolded SOD1. GAPDH and β-actin (Actin) were used for a loading control in Triton X-100-soluble and -insoluble fractions, respectively. c Quantification of 1% Triton X-100 soluble SOD1, insoluble SOD1_mono, insoluble SOD1_HMW, soluble misfolded SOD1, and insoluble misfolded SOD1. Values are mean ± s.e.m. (n = 4) in an arbitrary unit relative to H46R mice at 16 weeks of age. Signal intensities were normalized by the levels of GAPDH (soluble fractions) and Actin (insoluble fractions). Statistical significance was evaluated by two-way ANOVA with Bonferroni’s post hoc test (comparisons between different genotypes in the same age; *p < 0.05, **p < 0.01)